Oligonucleotide Method Validation

[jslindemann]

I understand that most method validations are written from the point of view of pharmaceutical research, e.g. quantitation of drug from plasma. However, my application of LC/MS is simply for purity and identity of a given synthesized piece of DNA (from 8 to 40-mers). We have a good LC/MS method, but it needs to be validated. Does anyone have any experience with validating this type of method, or do you know of any resources that might help?

[tom jupille]

Validation consists of demonstrating that a method does what it is purports to do. What you have to do to validate the method depends on who wants to know (and how much convincing they need). In a regulated environment (e.g., pharmaceuticals), "who wants to know" is the regulatory agencies, and "how much convincing they need" is spelled out in detail.

So "the ball is in your court".

For identity: Run a check sample that contains a known standard of your oligo plus all the other stuff it could possibly be and demonstrate that you can separate your product from the rest.

For purity: How accurately and precisely do you need to determine that? Once you have that answer, you can adapt the pharma validation procedures as necessary.